Meiyuan Qian, Yuefeng Tan, Bo Xu
Isicelo se-R&D Centre
Intshayelelo
I-Taxus (i-Taxus chinensis okanye i-yew yaseTshayina) sisityalo sasendle esikhuselwe lilizwe.Sisityalo esinqabileyo nesisengozini yokuphela esishiywe ngasemva yiQuaternary glaciers.Ikwakuphela kwesityalo samayeza endalo ehlabathini.I-Taxus isasazwe kummandla opholileyo we-hemisphere esemantla ukuya kumbindi we-subtropiki, malunga neentlobo ezili-11 zehlabathi.Kukho iintlobo ezi-4 kunye ne-1 ehlukeneyo e-China, oko kukuthi i-Northeast Taxus, i-Yunnan Taxus, i-Taxus, i-Tibetan Taxus kunye ne-Southern Taxus.Ezi ntlobo zintlanu zisasazwa kuMzantsi-ntshona weTshayina, eMzantsi China, kuMbindi weTshayina, eMpuma yeChina, kuMntla-ntshona weTshayina, kuMntla-mpuma weTshayina naseTaiwan.Izityalo ze-Taxus ziqulethe iindidi ezininzi zeekhemikhali, kuquka i-taxane, i-flavonoids, i-lignans, i-steroids, i-phenolic acids, i-sesquiterpenes kunye ne-glycosides.Ichiza elidumileyo lokulwa nethumba iTaxol (okanye iPaclitaxel) luhlobo lwerhafu.I-Taxol ineendlela ezizodwa zokulwa nomhlaza.I-Taxol “ingakhenkceza” ii-microtubules ngokudibanisa nazo kwaye ithintele ii-microtubules ekwahluleni iichromosomes ngexesha lokwahlulahlulwa kweeseli, nto leyo ekhokelela ekufeni kweeseli ezahlulayo, ngakumbi iiseli zomhlaza ezanda ngokukhawuleza[1].Ngaphezu koko, ngokuvula i-macrophages, i-Taxol ibangela ukuhla kwe-TNF-α (i-tumor necrosis factor) receptors kunye nokukhululwa kwe-TNF-α, ngaloo ndlela ibulala okanye inqanda iiseli ze-tumor [2].Ngaphezu koko, i-Taxol inokubangela i-apoptosis ngokusebenza kwindlela ye-apoptotic receptor ephakathi kwe-Fas/FasL okanye isebenze inkqubo ye-cysteine protease[3].Ngenxa yesiphumo se-anticancer ekujoliswe kuyo ngokuphindaphindiweyo, i-Taxol isetyenziswa ngokubanzi kunyango lomhlaza wesibeleko, umhlaza wamabele, umhlaza wemiphunga ongeyonxalenye encinci yeseli (NSCLC), umhlaza wesisu, umhlaza wommizo, umhlaza wesinye, umhlaza wesinyi, umhlaza wedlala lobudoda, imelanoma embi, intloko kunye nentamo. umhlaza, njalo njalo[4].Ngokukodwa kumhlaza wamabele ophambili kunye nomhlaza we-ovarian ophambili, iTaxol inesiphumo esibalaseleyo sonyango, ke ngoko yaziwa ngokuba "ngumgca wokugqibela wokuzikhusela kunyango lomhlaza".
I-Taxol lelona chiza lidumileyo lokulwa nomhlaza kwimarike yamazwe ngamazwe kwiminyaka yakutshanje kwaye lithathwa njengelona chiza lisebenzayo lokulwa nomhlaza ebantwini kule minyaka ingama-20 izayo.Kwiminyaka yakutshanje, ngokukhula okugqabhukileyo kwabemi kunye neziganeko zomhlaza, imfuno yeTaxol nayo inyuke kakhulu.Okwangoku, iTaxol efunekayo kuphando lwezonyango okanye lwezenzululwazi ikakhulu itsalwa ngqo kwiTaxus.Ngelishwa, umxholo weTaxol kwizityalo uphantsi kakhulu.Umzekelo, umxholo weTaxol yi-0.069% kuphela kwixolo le-Taxus brevifolia, elithathwa ngokubanzi njengowona mxholo uphezulu.Ukukhutshwa kwe-1 g yeTaxol, kufuna malunga ne-13.6 kg ye-Taxus bark.Ngokusekelwe kolu qikelelo, kuthatha 3 – 12 imithi yeTaxus engaphezu kweminyaka eli-100 ubudala ukunyanga isigulana somhlaza wesibeleko.Ngenxa yoko, inani elikhulu lemithi ye-Taxus liye lagawulwa, nto leyo ekhokelela ekuphelisweni kolu didi luxabisekileyo.Ukongeza, i-Taxus ihlupheke kakhulu kwimithombo kwaye ikhula kade, nto leyo eyenza kube nzima uphuhliso oluthe kratya kunye nokusetyenziswa kweTaxol.
Okwangoku, ukudityaniswa okupheleleyo kweTaxol kugqitywe ngempumelelo.Nangona kunjalo, indlela yayo yokwenziwa intsonkothe kakhulu kwaye ineendleko eziphezulu, iyenza ingabinamsebenzi wamashishini.Indlela ye-semi-synthetic ye-Taxol ngoku ikhulile kwaye ithathwa njengendlela esebenzayo yokwandisa umthombo we-Taxol ukongeza kutyalo olwenziweyo.Ngokufutshane, kwi-semi-synthesis yeTaxol, i-Taxol precursor ikhompawundi eninzi kakhulu kwizityalo ze-Taxus iyakhutshelwa emva koko iguqulelwe kwi-Taxol nge-chemical synthesis.Umxholo we-10-deacetylbaccatin Ⅲ kwiinaliti ze-Taxus baccata zinokufikelela kwi-0.1%.Kwaye iinaliti zilula ukuhlaziya ngokuthelekisa namaxolo.Ke ngoko, i-semi-synthesis yeTaxol esekwe kwi-10-deacetylbaccatin Ⅲ itsala ingqalelo engakumbi nangakumbi kubaphandi[5] (njengoko kubonisiwe kuMfanekiso 1).
Umzobo 1. Indlela yesemi-synthetic yeTaxol esekwe kwi-10-deacetylbaccatin Ⅲ.
Kwesi sithuba, isicatshulwa sesityalo se-Taxus sahlanjululwa nge-flash preparative liquid chromatography system SepaBean™ umatshini odityaniswe ne-SepaFlash C18 i-reversed-phase-phase (RP) flash cartridges eveliswe yiSantai Technologies.Imveliso ekujoliswe kuyo ihlangabezana neemfuno zococeko yafunyanwa kwaye ingasetyenziselwa uphando lwezenzululwazi olulandelayo, inikezela ngesisombululo esineendleko zokuhlanjululwa ngokukhawuleza kolu hlobo lweemveliso zendalo.
| Isixhobo | Umatshini weSepaBean™ | |
| Ikhatriji | 12 g SepaFlash C18 RP flash cartridge (silica spherical, 20 - 45μm, 100 Å, inombolo Order: SW-5222-012-SP) | |
| Ubude bamaza | 254 nm (ukubona), 280 nm (ukubeka iliso) | |
| Isigaba esiphathwayo | Isinyibilikisi A: Amanzi | |
| I-Solvent B: I-Methanol | ||
| Umgangatho wohambo | 15 mL/min | |
| Ukulayisha isampulu | I-20 mg isampuli eluhlaza echithwe kwi-1 mL DMSO | |
| Umgangatho | Ixesha (min) | Isinyibilikisi B (%) |
| 0 | 10 | |
| 5 | 10 | |
| 7 | 28 | |
| 14 | 28 | |
| 16 | 40 | |
| 20 | 60 | |
| 27 | 60 | |
| 30 | 72 | |
| 40 | 72 | |
| 43 | 100 | |
| 45 | 100 | |
Iziphumo neengxoxo
Ikhromatogram ekhanyayo yesicatshulwa ekrwada esivela kwi-Taxus ibonisiwe kuMfanekiso 2. Ngokuhlalutya ichromatogram, imveliso ekujoliswe kuyo kunye nokungcola okuphunyeziweyo kokwahlulwa kwesiseko.Ngaphaya koko, ukuphinda kuveliswe kakuhle kwaphunyezwa ngeenaliti ezininzi zeesampulu (idatha ayiboniswanga).Kuya kuthatha malunga neeyure ze-4 ukugqiba ukwahlukana kwindlela ye-chromatography ye-manual kunye neentsika zeglasi.Ukuthelekisa kunye nendlela ye-chromatography ye-manual manual, indlela yokucoca ngokuzenzekelayo kule post ifuna kuphela imizuzu ye-44 ukugqiba umsebenzi wokucoca (njengoko kuboniswe kuMfanekiso 3).Ngaphezulu kwe-80% yexesha kunye nesixa esikhulu se-solvent sinokugcinwa ngokuthatha indlela ezenzekelayo, enokunciphisa ngokufanelekileyo iindleko kunye nokuphucula kakhulu ukusebenza kakuhle.
Umzobo 2. Ikhromatogram ekhanyayo yesicatshulwa esikrwada esivela kwi-Taxus.
Umzobo 3. Ukuthelekiswa kwendlela ye-chromatography ye-manual kunye nendlela yokucoca ngokuzenzekelayo.
Ukuqukumbela, ukudibanisa i-SepaFlash C18 RP flash cartridges kunye nomatshini we-SepaBean™ unokubonelela ngesisombululo esikhawulezayo nesisebenzayo sokucocwa ngokukhawuleza kweemveliso zendalo ezifana ne-Taxus extract.
Iimbekiselo
1. I-Alushin GM, i-Lander GC, i-Kellogg EH, i-Zhang R, i-Baker D kunye ne-Nogales E. Izakhiwo ze-microtubule eziphakamileyo zityhila ukuguqulwa kwesakhiwo kwi-αβ-tubulin phezu kwe-GTP hydrolysis.Iseli, 2014, 157 (5), 1117-1129.
2. Burkhart CA, Berman JW, Swindell CS kunye neHorwitz SB.Ubudlelwane phakathi koLwakhiwo lweTaxol kunye nezinye iiRhafu kwi-Induction of Tumor Necrosis Factor-α Ukubonakaliswa kweGene kunye neCytotoxicity.Uphando lweCancer, 1994, 54 (22), 5779-5782.
3. Ipaki SJ, Wu CH, Gordon JD, Zhong X, Emami A kunye neSafa AR.I-Taxol yenza i-Caspase-10-exhomekeke kwi-Apoptosis, J. Biol.Chem., 2004, 279, 51057-51067.
4. I-Paclitaxel.I-American Society of Health-System Pharmacists.[NgoJanuwari 2, 2015]
5. UBruce Ganem noRoland R. Franke.I-Paclitaxel esuka kwi-Primary Taxanes: Umbono kwi-Creative Invention kwi-Organozirconium Chemistry.J. Org.Chem., 2007, 72 (11), 3981-3987.
Kukho uluhlu lwee-cartridges ze-SepaFlash C18 RP ezineenkcazo ezahlukeneyo ezivela kwi-Santai Technology (njengoko kuboniswe kwiThebhile 2).
| Inombolo yento | Ubungakanani beKholam | Umgangatho wohambo (mL/min) | Uxinzelelo olukhulu (psi/bar) |
| SW-5222-004-SP | 5.4 g | 5-15 | 400/27.5 |
| I-SW-5222-012-SP | 20 g | 10-25 | 400/27.5 |
| SW-5222-025-SP | 33 g | 10-25 | 400/27.5 |
| SW-5222-040-SP | 48 g | 15-30 | 400/27.5 |
| SW-5222-080-SP | 105 g | 25-50 | 350/24.0 |
| SW-5222-120-SP | 155 g | 30-60 | 300/20.7 |
| I-SW-5222-220-SP | 300 g | 40-80 | 300/20.7 |
| SW-5222-330-SP | 420 g | 40-80 | 250/17.2 |
Itheyibhile 2. SepaFlash C18 RP flash cartridges.
Izinto zokupakisha: Umgangatho ophezulu we-spherical C18-bonded silica, 20 - 45 μm, 100 Å
Ngolwazi oluthe vetshe ngeenkcukacha zomatshini weSepaBean™, okanye ulwazi lokuodola kwiikhatriji zeSepaFlash, nceda undwendwele iwebhusayithi yethu.
Ixesha lokuthumela: Sep-20-2018